Baoli YAO (Xi'an Institute of Optics and Precision Mechanics, CAS)
“100 fps/2048×2048 pixels/16-bits gray scale Structured Illumination Microscopy with DMD-based LED illumination”
|Title:||Fast Structured Illumination Microscopy|
|Author:||Baoli Yao, Ming Lei, Dan Dan, Xing Zhou, Jia Qian, Yanlong Yang, Shaohui Yan, Junwei Min, Xianghua Yu, Wei Zhao|
|Affiliation:||Professor, State Key Laboratory of Transient Optics and Photonics, Xi'an Institute of Optics and Precision Mechanics, Chinese Academy of Sciences|
Images captured by D-SIM. (a) BPAE cells (2D Super- resolution image), (b) DMD chip (3D image), (c) Mixed pollen grains (3D image), (d) Mite (3D image).
|Super-resolution three-dimensional optical microscopy has incomparable advantages over other high-resolution microscopic technologies, such as electron microscopy and atomic force microscopy, in the study of biological molecules, pathways and events in live cells and tissues. In this presentation, we report a strategy for fast super-resolution and 3D imaging based on the structured illumination microscopy (SIM), which is a wide-field technique to obtain either super-resolution or optical sectioning images comparable to the confocal laser scanning microscopy (CLSM). We built a DMD-based LED illumination SIM system (D-SIM), which has a fast fringe-shifting speed due to using the DMD device and a fast data acquisition rate by employing an ultra-sensitive sCMOS camera with its full potentiality of 100 fps at 2048×2048 pixels resolution and 16 bits gray scale. We developed a fast and robust algorithm for SIM super-resolution image reconstruction, and a modified algorithm for SIM optical sectioning image reconstruction based on Hilbert-transform that requires only two arbitrary phase-shifting raw images. In the super-resolution operation mode, the system can capture and display 2D super-resolution images in real time (~30fps). In the 3D optical sectioning operation mode, the system is able to reach a data acquisition rate of 418Mpixels/second, which is almost 100 times faster than the single focal spot CLSM microscopy under the same condition. The field of view can be extended by moving the sample stage transversally to obtain partially overlapped multiple images. An automatic imaging register algorithm is implemented to stitch those overlapped images in 2D or 3D spaces. The D-SIM is a good integration of cost-effective, ease of multi-wavelength switchable, speckle-noise-free, fast and multi-functional microscopic system.|
|Biographical Sketch:||Prof. Baoli Yao obtained his Ph.D. degree at Xi'an Institute of Optics and Precision Mechanics, CAS in 1997, and pursued the postdoctoral work in Technical University of Munich, Germany during 1997-1998. He is currently the deputy director of State Key Laboratory of Transient Optics and Photonics, China. His research areas include super-resolution optical microscopy, digital holographic microscopy, optical micro-manipulation and micro-fabrication, optical data storage and information processing. He invented the DMD-based LED-illumination structured illumination microscope (SIM), and got the 90nm lateral resolution. He designed and developed an optical tweezers system, which has been successfully commercialized and exported to Canada. He predicted theoretically the improvement of axial optical trapping efficiency of micro-particles by radially polarized beams. He proposed a new approach to generation of multiple isotropic super-diffraction-limit focused spots and the dynamic shifting method. He has authored more than 180 papers in peer-reviewed journals and 3 book’s chapters, and owned 17 approved Chinese invention patents. He obtained the High-Speed Imaging Award of Japan in 2015.|